Heterocyclic GSK-3 Allosteric Modulators

ABSTRACT

The present invention relates to heterocyclic substituted quinoline derivatives as allosteric inhibitors of the glycogen synthase kinase-3 (GSK-3) enzyme. Therefore, these compounds are useful for the manufacturing of a medicament designed for the treatment and/or prevention of diseases wherein GSK-3 is involved, such as neurodegenerative diseases, inflammatory diseases, cancer, diabetes, and to promote various regenerative processes.

The present invention relates to heterocyclic substituted quinolinederivatives as allosteric inhibitors of the glycogen synthase kinase-3(GSK-3) enzyme. Therefore, these compounds are useful for themanufacturing of a medicament designed for the treatment and/orprevention of diseases wherein GSK-3 is involved, such asneurodegenerative diseases, inflammatory diseases, cancer and diabetes,as well as to promote various regenerative processes. Therefore, theinvention may be included within the field of pharmaceutical chemistry.

STATE OF THE ART

Glycogen synthase kinase-3 (GSK-3) is an enzyme of the kinase familywhich catalyses the phosphorylation of serine or threonine residues invarious substrates. It was originally discovered due to its role inglycogen biosynthesis, after which it is named [Rylatt, D. B.; Aitken,A.; Bilham, T.; Condon, G. D.; Embi, N.; Cohen, P. Glycogen SynthaseKinase 3 from rabbit skeletal muscle. Separation fromcyclic-AMP-dependent protein kinase and, phosphorylase kinase. Eur JBiochem. 1980 107, 519-527]. This enzyme plays a key role in severalcell signalling pathways, including Wnt pathways, the cell divisioncycle, inflammation, cell proliferation, DNA damage response, cell deathand survival, and neuronal differentiation, amongst others [Phukan, S.;Babu, V. S.; Kannoji, A.; Hariharan, R.; Balaji, V. N. GSK3beta: role intherapeutic landscape and development of modulators. Br. J. Pharmacol.2010, 160, 1-19]. Recently, it has been demonstrated that anoverexpression/overactivation of GSK-3 is sufficient to induce neuronalcell death, and is related to various pathologies, such as bipolardisorders, neurodegenerative diseases, especially Alzheimer's disease,type II diabetes and chronic inflammatory diseases [Kannoji, A., Phukan,S., Sudher, V., Balaji, V. N. “GSK3beta: a master switch and a promisingtarget”. Expert Opin Ther Targets. 2008, 12, 1443-1455].

In recent years, numerous GSK-3 inhibitors have been synthesised, whichare promising molecules for the treatment of various diseases, such asdiabetes, cancer and neurodegenerative diseases [Martinez, A.Preclinical efficacy on GSK-3 inhibitors: towards a future generation ofpowerful drugs. Med. Res. Rev. 2008, 28, 773-796]. However, given thatthe human kinome is composed of more than 500 kinases with a highidentity at the catalytic site, i.e. the ATP-binding site, the discoveryand/or design of specific inhibitors of a given kinase is an openchallenge. One of the possibilities of increasing the selectivity ofkinases lies in the design of allosteric modulators [Eglen, R.; Reisine,T. Drug discovery and the human kinome: recent trends. Pharmacol. Ther.2011, 130, 144-156]. These compounds generally bind to unique, specificregions of the kinase, inducing conformational changes, and are veryuseful when resistances caused by the inhibitors that compete withadenosine triphosphate (ATP) appear [McInnes, C.; Fischer, P. M.Strategies for the design of potent and selective kinase inhibitors.Curr. Pharm. Des. 2005, 11, 1845-1863]. Moreover, allosteric modulatorsprovide mild modulation of the corresponding enzyme, which isparticularly important in the inhibition of GSK-3 [Martinez, A., Gil,C., Perez, D. I. Glycogen synthase kinase 3 inhibitors in the nexthorizon for Alzheimer's disease treatment. Int. J. Alzheimer's Dis.2011, doi: 10.406/2011/280502]. In this case, only the aberrantoverexpression/overactivation of GSK-3 must be inhibited in order totreat the pathologies wherein this kinase is involved.

DETAILED DESCRIPTION OF THE INVENTION

The present invention presents a family of compounds, and the method forobtaining them, that have the capacity to inhibit the GSK-3 enzyme inthe micromolar range. The kinetic studies of GSK-3 inhibition presentedhere show that these compounds are allosteric GSK-3 inhibitors. On theother hand, these compounds are capable of reducing inflammation in thecellular models presented herein.

A first aspect of the present invention relates to the use of a compoundwith the formula (I)

or the salts, solvates or stereoisomers thereof,

where R₁ is selected from H, an optionally substituted C₁-C₅ alkylgroup, an optionally substituted C₂-C₅ alkenyl group, R₂ is anoptionally substituted C₅-C₁₅ alkyl group, R₃ is selected from H,halogen, an optionally substituted C₁-C₅ alkyl group or an optionallysubstituted C₁-C₅ —(O)-alkyl group, n is a value between 1 and 4, R₄, R₅and R₆ are independently selected from H or an optionally substitutedC₁-C₅ alkyl group, provided that, when R₁ is ethyl or H and R₃, R₄ , R₅and R₆ are H, R₂ cannot be heptyl,

for the manufacturing of a medicament for the treatment and/orprevention of a disease selected from neurodegenerative diseases,inflammatory diseases, cancer, diabetes, or to promote regenerativeprocesses.

In the present invention, the term “alkyl” refers to linear or branchedhydrocarbonated chain radicals, with between 1 and 15 carbon atoms,which bind to the rest of the molecule by means of a single bond, forexample, propyl, ethyl, methyl, isopropyl, undecanoyl, heptadecanoyl,octadecanoyl, etc. These alkyl radicals may be optionally substituted inone or more positions by one or more groups, such as cycloalkyl,hydroxyl, amines, amides, oxo, cyano, halogens, aryl, etc.

In the present invention, the term “cycloalkyl” refers to cyclichydrocarbonated chain radicals, preferably with between 3 and 6 carbonatoms, and more preferably 3, totally or partially saturated, and formedby only carbon and hydrogen atoms, such as cyclopropyl, cyclopentyl orcyclohexyl, and which may be optionally substituted by one or moregroups, such as alkyl, halogens, hydroxyl, amines, amides, cyano, etc.

In the present invention, the term “alkenyl” refers to linear orbranched hydrocarbonated chain radicals, which contain one or moredouble carbon-carbon bonds, for example, vinyl, 1-propenyl, allyl,isoprenyl, 2-butenyl, 1,3-butadienyl, etc. The alkenyl radicals may beoptionally substituted by one or more groups, such as halogen, hydroxyl,carboxyl, cyano, carbonyl, acyl, amino, nitro, etc.

In the present invention, the term “aryl” refers to single or multiplearomatic rings, which have between 5 and 18 links wherefrom a proton hasbeen eliminated from the ring. Preferably, the aryl group has between 5and 7 carbon atoms. Aryl groups are, for example, without being limitedthereto, phenyl, naphthyl, diphenyl, indenyl, phenanthryl or anthracyl.The aryl radicals may be optionally substituted by one or moresubstituents, such as alkyl, hydroxyl, amines, amide, cyano, halogens,etc.

“Halogen” refers to fluorine, chlorine, bromine or iodine.

In a preferred embodiment of the present invention, R₄, R₅ and R₆ are H.

In another preferred embodiment of the present invention, R₁ is H or aC₁-C₃ alkyl group; more preferably, R₁ is H, optionally substitutedmethyl, ethyl or isobutyl. In a more preferred embodiment, R₁ is amethyl group substituted by a cycloalkyl, preferably cyclopropyl, or anaryl, preferably phenyl.

In another preferred embodiment of the present invention, R₂ is a C₇-C₁₁alkyl group; more preferably, R₂ is a C₇ alkyl group or a C₁₁ alkylgroup.

In the compound with the formula (I) of the invention, when R₃ is ahalogen group, it is preferably Br, Cl or F, and, more preferably, n is1.

In the compound with the formula (I) of the invention, when R₃ is H, nis preferably 4.

A more preferred embodiment relates to the use of a compound selectedfrom the following group:

-   -   4-Hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (3)    -   N′-Dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (4)    -   N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (6)    -   N′-Dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (13)    -   7-Chloro-4-hydroxy-N′-octanoyl-2-oxo-2-hydroquinoline-3-carbohydrazide        (14)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (15)    -   6-Fluoro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (17)    -   4-Hydroxy-1-(isoprenyl)-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (42)    -   N′-Dodecanoyl-4-hydroxy-1-(isoprenyl)-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (43)    -   N′-Dodecanoyl-4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (44)    -   1-(Cyclopropylmethyl)-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (45)    -   1-Benzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (46)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (47)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (48)    -   6-Bromo-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (49)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (50)    -   6-Bromo-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (51)    -   6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (52)    -   6-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (53)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (54)    -   6-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (55)    -   6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (56)    -   7-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (57)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (58)    -   7-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (59)    -   7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (60)    -   1-Benzyl-7-chloro-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (61)    -   1-Benzyl-7-chloro-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (62)    -   N′,1-Dibenzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (63)

Another preferred embodiment relates to the use of a compound selectedfrom the following group:

-   -   N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (4)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroguinoline-3-carbohydrazide        (15)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroguinoline-3-carbohydrazide        (47)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroguinoline-3-carbohydrazide        (48)    -   6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (52)    -   6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (56)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (58)    -   7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (60)

or the salts, solvates or stereoisomers thereof, for the manufacturingof a medicament designed for the treatment and/or prevention of adisease selected from neurodegenerative diseases, inflammatory diseases,cancer, diabetes, or to promote regenerative processes.

The compounds of the present invention, both those with the generalformula (I) and those with the general formula (II), are allostericGSK-3 inhibitors; therefore, these compounds are used for the treatmentand/or prevention of diseases wherein this enzyme is involved.Allosteric inhibitors produce milder modulation of enzymes thancompetitive inhibitors, with IC50 values in the micromolar range. TheGSK-3 enzyme is involved in multiple cell signalling pathways; for thisreason, inhibitors of this enzyme aimed at a specific pathway related toa specific pathology end up affecting other pathways, thereby generatingundesirable secondary effects. Allosteric modulation of GSK-3 entails alower impact on those pathways not related to the disease to be treated,which would minimise secondary effects in a patient.

The neurodegenerative diseases may be selected from the list thatcomprises, without being limited thereto, Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, cerebral ischaemia,post-encephalitic Parkinsonisms, dystonias, Tourette syndrome, periodiclimb movement pathologies, restless legs syndrome, attention deficithyperactivity disorders, Huntington's disease, progressive supranuclearpalsy, Pick's disease, fronto-temporal dementia or neuromusculardiseases.

The inflammatory diseases may be selected, without being limitedthereto, from Crohn's disease, ulcerative colitis, rheumatoid arthritis,atherosclerosis, vasculitis or multiple sclerosis.

The cancer may be selected, without being limited thereto, fromglioblastoma, leukaemias, lymphomas, or lung, breast, prostate or coloncancer.

The diabetes may be non-insulin-dependent type II diabetes.

Preferably, in the regenerative process to be promoted by the use ofcompounds with the formula (I), the differentiation of the followingstem cells is involved: nervous system stem cells, haematopoietic systemstem cells, skeletal system stem cells or myocardial stem cells.

Another aspect of the present invention relates to a compound with theformula (II):

or the salts, solvates or stereoisomers thereof,

where R₁ is selected from H or an optionally substituted C₁-C₅ alkylgroup or an optionally substituted C₂-C₅ alkenyl group, R₂ is anoptionally substituted C₅-C₁₅ alkyl group, R₃ is a halogen, n is a valuebetween 1 and 4, R₄, R₅ and R₆ are independently selected from H or anoptionally substituted C₁-C₅ alkyl group.

In a preferred embodiment of the compounds of the present invention, R₄,R₅ and R₆ are H.

In another preferred embodiment of the compounds of the presentinvention, R₁ is H or a C₁-C₃ alkyl group, preferably optionallysubstituted methyl, ethyl or isobutyl.

In a more preferred embodiment, R₁ is a methyl group substituted by acycloalkyl, preferably cyclopropyl, or an aryl, preferably phenyl.

In another more preferred embodiment, R₁ is H.

In another preferred embodiment of the compounds of the presentinvention, R₂ is a C₇-C₁₁ alkyl group; more preferably, R₂ is a C₇ alkylgroup or a C₁₁ alkyl group.

In another preferred embodiment, R₃ is Br, Cl or F, and n is preferably1.

In another preferred embodiment, R₃ is H and n is preferably 1.

A more preferred embodiment of the compounds with the formula (II)comprises the following compounds:

-   -   7-Chloro-4-hydroxy-N′-octanoyl-2-oxo-2-hydroquinoline-3-carbohydrazide        (14)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (15)    -   6-Fluoro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (17)    -   4-Hydroxy-1(isoprenyl)-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (42)    -   N′-Dodecanoyl-4-hydroxy-1-(isoprenyl)-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (43)    -   N′-Dodecanoyl-4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (44)    -   1-(Cyclopropylmethyl)-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (45)    -   1-Benzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (46)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (47)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (48)    -   6-Bromo-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (49)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (50)    -   6-Bromo-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (51)    -   6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (52)    -   6-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (53)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (54)    -   6-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (55)    -   6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (56)    -   7-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (57)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (58)    -   7-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (59)    -   7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (60)    -   1-Benzyl-7-chloro-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (61)    -   1-Benzyl-7-chloro-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (62)    -   N′,1-Dibenzyl-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (63)

Another preferred embodiment relates to a compound selected from thefollowing group:

-   -   N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (6)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroguinoline-3-carbohydrazide        (15)    -   6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroguinoline-3-carbohydrazide        (47)    -   6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide        (48)    -   6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (52)    -   6-chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (56)    -   7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (58)    -   7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide        (60)

A third aspect of the present invention relates to the use of a compoundwith the formula (II), as described above, for the manufacturing of amedicament.

A fourth aspect of the present invention relates to a pharmaceuticalcomposition that comprises a compound with the formula (II), asdescribed above, jointly with a pharmaceutically adequate vehicle. In apreferred embodiment, this composition further comprises another knownactive principle, used for the treatment and/or prevention of any of theaforementioned diseases.

The compounds of the present invention represented by the formula (I) orthe formula (II), and, more specifically, the specific compoundspertaining to this previously described general formula may includeisomers, depending on the presence of multiple bonds (for example, Z,E), including optical isomers or enantiomers, depending on the presenceof chiral centres. The individual isomers, enantiomers ordiastereoisomers, and the mixtures thereof, fall within the scope of thepresent invention. The individual enantiomers or diastereoisomers, andthe mixtures thereof, may be separated by means of conventionaltechniques.

The compounds of the invention may be in crystalline form as freecompounds or as solvates, and both forms are intended to be includedwithin the scope of the present invention. In this regard, the term“solvate”, as used herein, includes both pharmaceutically acceptablesolvates, i.e. solvates of the compound with the formula (I) that may beused in the preparation of a medicament, and pharmaceuticallyunacceptable solvates, which may be useful in the preparation ofpharmaceutically acceptable solvates or salts. The nature of thepharmaceutically acceptable solvate is not critical, provided that it ispharmaceutically acceptable. In a particular embodiment, the solvate isa hydrate. The solvates may be obtained by conventional solvationmethods that are well-known to persons skilled in the art.

The compounds with the formula (I) or the formula (II) designed fortherapeutic use are prepared in solid form or aqueous suspension, in apharmaceutically acceptable diluent. These preparations may beadministered by any appropriate administration route, for which reasonsaid preparation will be formulated in the adequate pharmaceutical formfor the selected administration route. In a particular embodiment,administration of the compound with the formula (I) provided by thisinvention is performed by oral, topical, rectal or parenteral route(including subcutaneous, intraperitoneal, intradermal, intramuscular,intravenous route, etc.). A review of the different pharmaceutical formsfor the administration of medicaments and the necessary excipients toobtain them may be found, for example, in “Tratado de FarmaciaGalénica”, C. Faulí i Trillo, 1993, Luzán 5, S. A. Ediciones, Madrid, orother habitual or similar ones in the Spanish Pharmacopeia and in theUnited States.

The compounds described in the present invention, their pharmaceuticallyacceptable salts and solvates, and the pharmaceutical compositionscontaining them may be used jointly with other, additional drugs, toprovide combined therapy. Said additional drugs may be a part of thesame pharmaceutical composition or, alternatively, may be provided inthe form of a separate composition for simultaneous or non-simultaneousadministration with the pharmaceutical composition comprising a compoundwith the formula (I) or the formula (II), or a pharmaceuticallyacceptable salt, stereoisomer or solvate thereof.

Except as otherwise specified, the compounds of the invention alsoinclude compounds that differ only in the presence of one or moreisotope-enriched atoms. For example, compounds that have said structure,except for the substitution of a hydrogen atom by a deuterium or atritium atom, or the substitution of a carbon atom by an enriched carbonin ¹³C or ¹⁴C, or a nitrogen atom enriched in ¹⁵N, are included withinthe scope of this invention.

A final aspect of the present invention relates to a process forobtaining a compound with the formula (II), which comprises thefollowing steps:

-   -   1) Reaction of a resin with a compound with the formula (III)

where R₇ and R₈ are identical or different C₁-C₅ groups alkyl.

-   -   2) Reaction of the product obtained in the preceding step with a        product with the formula (IV):

where R₃ is defined as above.

-   -   3) Reaction of the product obtained in the preceding step with a        compound with the formula (V):

where R₉ is a C₁-C₅ alkyl group.

-   -   4) Cyclisation of the product obtained in the preceding step.    -   5) Reaction of the product obtained in the preceding step with a        compound with the formula (VI):

where R₂ is defined as above.

In a preferred embodiment of the process of the invention, R₇ and R₈ areindependently a C₁-C₃alkyl group and, more preferably, a methyl group.In a preferred embodiment of the process of the invention, R₉ is aC₁-C₃, alkyl group and, more preferably, it is an ethyl group.

Throughout the description and the claims, the word “comprises” and thevariants thereof are not intended to exclude other technicalcharacteristics, additives, components or steps. For persons skilled inthe art, other objects, advantages and characteristics of the inventionwill arise partly from the description and partly from the practise ofthe invention. The following examples and figures are provided forillustrative purposes, and are not intended to limit the scope of thisinvention.

FIGURES

FIG. 1 shows the double-reciprocal plot. The ATP concentrations werevaried from 1 to 50 μM, whereas the concentration of the GS-2 peptidewas kept constant at 12.5 μM. The concentrations of inhibitor (compound6) used are shown in the plot.

FIG. 2 shows the double-reciprocal plot. The GS-2 concentrations werevaried from 15.5 to 100 μM, whereas the ATP concentration was keptconstant at 1 μM. The concentrations of inhibitor (compound 6) used areshown in the plot.

FIG. 3 shows the decrease in inflammatory activity in cell cultures inthe presence of variable concentrations of compound 6.

EXAMPLES

Below, we will illustrate the invention by means of assays performed bythe inventors, which demonstrate the specificity and effectiveness ofthe compounds of the invention.

Compounds 1-17 were synthesised according to the synthetic pathwaysshown in diagrams 1 and 2.

General Synthesis Process for Compounds 1 and 2

The corresponding anhydride (1 equiv) is dissolved in 40 ml of DMF, andthe sodium hydride (1.2 equiv) and diethyl malonate (1.2 equiv) areadded. Subsequently, it is heated to 85° C. for 6 h. After this time haselapsed, the mixture is allowed to cool and acidified with a 1 M HClsolution. When it reaches an acid medium, it precipitates a white solidwhich, after being filtered, is the pure final compound.

Ethyl 4-Hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (1).Yield 56%. ¹H NMR (300 MHz, CDCl₃) δ 8.12 (d, J=8.0 Hz, 1H), 7.64 (t,J=7.9 Hz, 1H), 7.36-7.11 (m, 3H), 4.48 (q, J=7.1 Hz, 2H), 3.61 (s, 3H),1.99 (sa, 1H), 1.46 (t, J=7.1 Hz, 3H). ¹³C NMR (75 MHz, CDCl₃) δ 172.8,171.8, 159.7, 141.4, 134.5, 125.9, 122.0, 115.0, 114.3, 98.1, 62.4,29.35, 14.44. M.p.=95° C.-96° C. HPLC: Purity>99%. MS (ES): m/z=248,201, 133. Elementary Analysis (C₁₃H₁₃NO₄) Calculated: C, 63.15%; H,5.30%; N, 5.67%. Found: C, 63.31%; H, 5.08%; N, 5.77%.

Ethyl 1-Ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate (2).Yield 56%. ¹H NMR (300 MHz, CDCl₃) δ 14.22 (s, 1H), 8.20 (dd, J=8.1, 1.5Hz, 1H), 7.68 (ddd, J=8.7, 7.2, 1.6 Hz, 1H), 7.39-7.16 (m, 2H), 4.51 (q,J=7.1 Hz, 2H), 4.31 (q, J=7.1 Hz, 2H), 1.49 (t, J=7.1 Hz, 3H), 1.34 (t,J=7.1 Hz, 3H). ¹³C NMR (75 MHz, CDCl₃) δ 172.5, 171.4, 159.0, 140.1,134.1, 125.7, 121.5, 114.9, 113.8, 97.7, 62.1, 37.0, 14.0, 12.5.M.p.=68° C.-69° C. HPLC: Purity 97%. MS (ES): m/z=263, 217, 187.Elementary analysis (C₁₄H₁₅NO₄) Calculated: C, 64.36%; H, 5.79%; N,5.36%. Found: C, 64.72%; H, 5.63%; N, 5.05%.

General Synthesis Process for Compounds 3-6

1 equivalent of 1 or 2, as specified in each case, the correspondinghydrazide (1 equiv) and 0.2 ml of DMF are placed in a flask, and heatedto 160° C. for 3 min. It is allowed to cool, MeOH is carefully added,and a white solid precipitates, which is filtered and purified by meansof MeOH washings.

4-Hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(3). Reagents: Compound 1 and octanoyl hydrazide.

Yield 99%. ¹H NMR (300 MHz, CDCl₃) δ 15.49 (s, 1H), 12.46 (d, J=5.6 Hz,1H), 8.28 (d, J=5.7 Hz, 1H), 8.18 (dd, J=8.1, 1.5 Hz, 1H), 7.71 (t,J=7.9 Hz, 1H), 7.44-7.27 (m, 2H), 3.69 (s, 3H), 2.44-2.20 (t, J=7.6 Hz,2H), 1.90-1.56 (m, 2H), 1.45-1.18 (m, 8H), 0.88 (t, J=6.8 Hz, 3H). ¹³CNMR (75 MHz, CDCl₃) δ 172.4, 171.4, 166.9, 162.4, 140.2, 134.8, 126.2,129.9, 115.8, 115.1, 96.1, 34.4, 31.8, 31.3, 29.6, 29.4, 25.4, 22.7,14.1. M.p.=157° C.-158° C. HPLC: Purity>99%. MS (ES): m/z=360, 234.Elementary Analysis (C₁₉H₂₅N₃O₄) Calculated: C, 63.49%; H, 7.01%; N,11.69%. Found: C, 63.76%; H, 6.94%; N, 11.65%.

N′-Dodecanoyl-4-hydroxy-1-methyl-2-oxo-1.2-dihydroquinoline-3-carbohydrazide(4). Reagents: Compound 1 and dodecanoyl hydrazide.

Yield 93%. ¹H NMR (300 MHz, DMSO) δ 15.50 (s, 1H), 12.47 (d, J=5.6 Hz,1H), 8.20 (d, J=8.1 Hz, 1H), 8.16 (d, J=5.9 Hz, 1H), 7.72 (t, J=8.0 Hz,1H), 7.42-7.29 (m, 1H), 3.70 (s, 3H), 2.32 (t, J=7.6 Hz, 2H), 1.77-1.66(m, 2H), 1.26 (s, 16H), 0.88 (t, J=6.6 Hz, 3H). ¹³C NMR (75 MHz, CDCl₃)δ 171.1, 169.0, 166.6, 161.9, 140.1, 134.1, 125.4, 122.5, 115.6, 114.3,96.2, 34.4, 31.8, 29.5, 29.4, 29.3, 29.1, 25.4, 22.6, 14.0. M.p.=219°C.-220° C. HPLC: Purity>99%. MS (ES): m/z=416, 234. Elementary Analysis(C₂₃H₃₃N₃O₄) Calculated: C, 66.48%; H, 8.00%; N, 10.11%. Found: C,64.76%; H, 7.79%; N, 10.27%.

N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(6). Reagents: Compound 2 and dodecanoyl hydrazide.

Yield 91%. ¹H NMR (300 MHz, DMSO) δ 16.22 (s, 1H), 12.05 (d, J=3.7 Hz,1H), 10.80 (d, J=3.7 Hz, 1H), 8.09 (d, J=8.0 Hz, 1H), 7.82 (t, J=7.1 Hz,1H), 7.68 (d, J=8.6 Hz, 1H), 7.38 (t, J=7.4 Hz, 1H), 4.31 (d, J=6.7 Hz,2H), 2.24 (m, 2H), 1.55 (m, 2H), 1.23 (m, 19H), 0.84 (m, 3H). ¹³C NMR(75 MHz, DMSO) δ 171.3, 170.3, 167.9, 161.5, 139.5, 135.2, 125.3, 123.2,115.8, 96.4, 37.6, 33.7, 31.9, 29.6, 29.5, 29.3, 29.2, 25.6, 22.7, 14.5,13.3. M.p.=133° C.-134° C. HPLC: Purity>99%. MS (ES): m/z=430, 248.Elementary Analysis (C₂₄H₃₅N₃O₄) Calculated: C, 67.11%; H, 8.21%; N,9.78%. Found: C, 67.40%; H, 8.31%; N, 9.88%.

General Synthesis Process for Compounds 12-17

The Merrifield resin (7) (1 equiv) is added on dry dimethylacetamide andthe mixture is bubbled with argon. 4-hydroxy-2,6-dimethoxybenzaldehyde(2.13 equiv) dissolved in dimethylacetamide and cesium carbonate (2.56equiv) are added. The vial is hermetically sealed and allowed to reactat 85° C. for 36 h. After conveniently washing resin 8, drydichloromethane, the corresponding methyl benzoate derivative (3 equiv)and sodium triacetoxyborohydride (3 equiv) are added. The mixture isleft under stirring at room temperature for 16 h. Following thecorresponding washings, dry dichloromethane, ethyl monomalonate (3equiv), phosphoryl oxychloride (3 equiv) and pyridine (6 equiv) areadded to resin 9, and it is left under stirring at room temperature for2 h. After the corresponding washings, dry THF and a 0.5 M solution ofKHMDS in toluene (10 equiv) are added to resin 10, and it is allowed toreact for 12 h at room temperature. Following the correspondingwashings, resin 11 is unanchored using a TFACH₂Cl₂ solution (50:50)under stirring for 1 h at room temperature. The mixture is filtered andthe filtrate is washed with water. The organic phase is dried with MgSO₄and the solvent is evaporated at reduced pressure. The product obtainedis made to react with the corresponding hydrazide (1 equiv) in DMF andheated to 160° C. for 3 min. It is allowed to cool, MeOH is added, and awhite solid precipitates, which is filtered and subsequently purified bymeans of MeOH washings.

N′-Dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide (13).Reagents: methyl anthranilate and dodecanoyl hydrazide.

Yield 98%. ¹H NMR (300 MHz, DMSO) δ 16.11 (s, 1H), 11.98 (s, 1H), 11.92(s, 1H), 10.63 (s, 1H), 7.98 (d, J=8.1 Hz, 1H), 7.71 (t, J=7.7 Hz, 1H),7.39 (d, J=8.3 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 2.23 (t, J=7.2 Hz, 2H),1.57 (d, J=6.6 Hz, 2H), 1.25 (s, 16H), 0.85 (t, J=6.2 Hz, 3H). ¹³C NMR(75 MHz, DMSO) δ 172.2, 170.0, 167.8, 162.3, 139.1, 134.5, 124.3, 122.9,116.3, 114.5, 99.4, 33.2, 31.6, 31.0, 29.3, 29.0, 28.8, 25.3, 22.4,14.3. M.p.=256° C.-257° C. HPLC: Purity>99%. MS (ES): m/z=402, 220.Elementary Analysis (C₂₂H₃₁N₃O₄) Calculated: C, 65.81%; H, 7.78%; N,10.47%. Found: C, 66.02%; H, 7.83%; N, 10.74%.

7-Chloro-4-hydroxy-N′-octanoyl-2-oxo-2-hydroquinoline-3-carbohydrazide(14). Reagents: methyl 2-amino-4-chlorobenzoate and octanoyl hydrazide.

Yield 86%. ¹H NMR (300 MHz, DMSO) δ 16.11 (sa, 1H), 12.09 (s, 1H), 11.79(s, 1H), 10.71 (d, J=3.1 Hz, 1H), 7.94 (d, J=5.0 Hz, 1H), 7.48-7.16 (m,2H), 2.87 (s, 2H), 2.71 (s, 2H), 2.49 (s, 2H), 2.21 (t, J=7.3 Hz, 2H),1.61-1.42 (m, 2H), 1.25 (s, 2H), 0.85 (t, J=6.6 Hz, 3H). M.p.=290°C.-291° C. HPLC: Purity 95%. MS (ES): m/z=380, 254, 222. ElementaryAnalysis (C₁₈H₂₂ClN₃O₄) Calculated: C, 56.92%; H, 5.84%; N, 11.06%.Found: C, 56.84%; H, 5.73%; N, 11.00%.7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide(15). Reagents: methyl 2-amino-4-chlorobenzoate and dodecanoylhydrazide.

Yield last reaction 92%. ¹H NMR (300 MHz, DMSO) δ 16.21 (s, 1H), 12.10(s, 1H), 11.80 (s, 1H), 10.71 (s, 1H), 7.98 (d, J=8.6 Hz, 1H), 7.42 (d,J=1.8 Hz, 1H), 7.31 (dd, J=8.7, 2.0 Hz, 1H), 2.87 (s, 2H), 2.71 (s, 2H),2.20 (t, J=7.4 Hz, 2H), 1.53 (s, 2H), 1.23 (s, 12H), 0.82 (d, J=6.7 Hz,3H). M.p.=291-292° C. HPLC: Purity 97%. MS (ES): m/z=436, 254, 222.Elementary Analysis (C₂₂H₃₀ClN₃O₄) Calculated: C, 60.42%; H, 6.94%; N,9.64%. Found: C, 60.42%; H, 6.71%; N, 9.58%.6-Fluoro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide(17). Reagents: methyl 2-amino-5-fluorobenzoate and dodecanoylhydrazide.

Yield 93%. ¹H NMR (400 MHz, DMSO) δ 11.86 (s, 1H), 10.42 (s, 1H), 7.64(d, J=6.9 Hz, 1H), 7.56 (td, J=8.7, 2.9 Hz, 1H), 7.42 (dd, J=9.2, 4.5Hz, 1H), 2.47 (m, 2H), 2.21 (t, J=7.3 Hz, 2H), 1.66-1.42 (m, 2H), 1.24(s, 14H), 0.83 (t, J=6.9 Hz, 3H). M.p.=178° C.-179° C. HPLC: Purity 95%.MS (ES): m/z=420, 238, 206. Elementary Analysis (C₂₂H₃₀FN₃O₄)Calculated: C, 62.99%; H, 7.21%; N, 10.02%. Found: C, 63.20%; H, 7.42%;N, 9.87%.

General Synthesis Process for Compounds 18-28

The corresponding isatoic anhydride (1 equiv) is dissolved in 15 ml ofanhydrous DMF and, subsequently, sodium hydride (1.5 equiv) is addedslowly at 0° C. After 30 minutes, the corresponding halogenatedderivative is added (1.5-3 equiv) and the reaction is left understirring at room temperature overnight. Subsequently, dichloromethane(15 ml) and water (15 ml) are added. The organic phase is separated, andthe aqueous phase is once again extracted with dichloromethane (15 ml).The organic phases are brought together and washed with a saturated NaClsolution (15 ml). The organic phase is dried on anhydrous magnesiumsulfate and filtered, and the solvent is evaporated under vacuum. Theresulting solid is recrystallised in dichloromethane/hexane, to producethe desired final product.

N-isoprenyl isatoic anhydride (18).

Reagents: Isatoic anhydride (6.13 mmols, 1 g); Sodium hydride (9.2mmols, 0.23 g); Isoprenyl bromide (12 mmols, 1.9 g). Yield: 0.95 g(67%), white solid, m.p.=119° C.-120° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 8.01 (d, J=1.6, 7.8 Hz, 1H), 7.85 (t,J=1.6, 7.3, 8.8 Hz, 1H), 7.32-7.26 (m, 2H), 5.19 (t, J=1.4, 6.4 Hz, 1H),4.64 (d, J=6.2 Hz, 2H), 1.81 (s, J=1.4 Hz, 3H), 1.70 (s, J=1.4 Hz, 3H);¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 159.9, 147.6, 141.3, 137.1, 136.4,129.6, 123.6, 118.2, 111.9, 42.7, 25.4, 18.1. HPLC: Purity>99%, r.t.=8.9min. ESI MS (m/z): 232 [M+H]⁺.

N-isobutyl isatoic anhydride (19).

Reagents: Isatoic anhydride (6.13 mmols, 1 g); Sodium hydride (9.2mmols, 0.22 g); isobutyl iodide (18 mmols, 3.4 g). Yield: 0.6 g (46%),white solid, m.p.=85° C.-86° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 8.10 (dd, J=1.7, 7.9 Hz, 1H), 7.68(ddd, J=1.6, 7.3, 8.4 Hz, 1H), 7.25-7.18 (m, 1H), 7.10 (d, J=8.5 Hz, 1H)3.93 (d, J=7.6 Hz, 2H), 2.25-2.10 (m, 1H), 1.03 (d, J=6.7 Hz, 6H); ¹³CNMR (75 MHz, CDCl₃) δ (ppm): 162.4, 158.7, 148.3, 141.8, 137.2, 131.1,124.0, 114.4, 112.0, 51.7, 26.9, 20.1, 19.9. HPLC: Purity>99%, r.t.=8.6min. ESI MS (m/z): 220 [M+H]⁺.

N-(cyclopropylmethyl)isatoic anhydride (20).

Reagents: Isatoic anhydride (4.9 mmols, 0.8 g); Sodium hydride (5.9mmols, 0.14 g); Cyclopropylmethyl bromide (5.9 mmols, 0.8 g). Yield:0.53 g (50%), light brown solid, m.p.=122° C.-123° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 8.11 (dd, J=1.6, 7.7 Hz, 1H),7.75-7.67 (m, 1H), 7.27-7.20 (m, 2H), 3.95 (d, J=7.0 Hz, 2H), 1.25-1.10(m, 1H), 0.54-0.40 (m, 4H); ¹³C NMR (75 MHz, CDCl₃) δ (ppm): 158.7,148.4, 141.7, 137.3, 131.0, 124.0, 114.3, 111.9, 49.2, 9.3, 4.1. HPLC:Purity>97%, r.t.=8.2 min. ESI MS (m/z): 218 [M+H]⁺.

N-benzyl isatoic anhydride (21).

Reagents: Isatoic anhydride (6.13 mmols, 1 g); Sodium hydride (9.2mmols, 0.22 g); Benzyl bromide (12.3 mmols, 1.4 g). Yield: 0.2 g (15%),white solid, m.p.=139° C.-140° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 8.10 (dd, J=1.6, 7.8 Hz, 1H), 7.56 (td,J=1.6, 7.3, 8.7 Hz, 1H), 7.35-7.10 (m, 6H), 7.05 (dd, J=8.5 Hz, 1H),5.24 (s, 2H); ¹³C NMR (75 MHz, CDCl₃) δ (ppm): 157.3, 147.4, 140.3,136.2, 133.4, 129.8, 128.1, 127.1, 125.7, 123.1, 113.7, 110.8, 47.5.HPLC: Purity>99%, r.t.=8.7 min. ESI MS (m/z): 254 [M+H]⁺.

6-bromo-N-methyl isatoic anhydride (22).

Reagents: 5-bromo isatoic anhydride (4.13 mmols, 1 g); Sodium hydride(6.2 mmols, 0.15 g); Methyl iodide (6.2 mmols, 0.9 g). Yield: 0.69 g(66%), white solid, m.p.=201° C.-202° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 8.06 (d, J=2.5 Hz, 1H), 7.64 (dd,J=2.5, 9.0 Hz, 1H), 7.08 (d, J=9.0 Hz, 1H), 2.92 (s, 3H); ^(—)C NMR (75MHz, CDCl₃) δ (ppm): 157.2, 147.4, 141.0, 140.0, 133.0, 116.9, 115.6,113.2, 32.0. HPLC: Purity>98%, r.t.=7.7 min. ESI MS (m/z): 256 [M+H]⁺.

6-bromo-N-ethyl isatoic anhydride (23).

Reagents: 5-bromo isatoic anhydride (8.26 mmols, 2 g); Sodium hydride(12.5 mmols, 0.3 g); Ethyl iodide (12.5 mmols, 1.9 g). Yield: 1.4 g(63%), white solid, m.p.=177° C.-178° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 8.21 (d, J=2.4 Hz, 1H), 7.78 (dd,J=2.4, 8.9 Hz, 1H), 7.02 (d, J=8.9 Hz, 1H), 4.05 (c, J=7.2 Hz, 2H), 1.31(t, J=7.2 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 157.3, 146.9,140.0, 139.0, 133.2, 116.6, 115.5, 113.4, 40.3, 12.0. HPLC: Purity>97%,r.t.=8.4 min. ESI MS (m/z): 270 [M+H]⁺.

6-chloro-N-methyl isatoic anhydride (24).

Reagents: 5-chloro isatoic anhydride (3.97 mmols, 0.78 g); Sodiumhydride (5.95 mmols, 0.14 g); Methyl iodide (11.4 mmols, 1.6 g). Yield:0.5 g (60%), white solid, m.p.=203° C.-204° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 8.05 (d, J=2.5 Hz, 1H), 7.66 (dd,J=2.5, 8.9 Hz, 1H), 7.09 (d, J=8.9 Hz, 1H), 3.52 (s, 3H); ^(—)C NMR (75MHz, CDCl₃) δ (ppm): 156.3, 146.4, 139.5, 136.2, 129.0, 114.4, 111.9.HPLC: Purity>98%, r.t.=7.3 min.

6-chloro-N-ethyl isatoic anhydride (25).

Reagents: 5-chloro isatoic anhydride (2.53 mmols, 0.5 g); Sodium hydride(3.8 mmols, 0.09 g); Ethyl iodide (3.8 mmols, 0.6 g). Yield: 0.4 g(70%), white solid, m.p.=150° C.-151° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 8.07 (d, J=2.5 Hz, 1H), 7.64 (dd,J=2.5, 8.9 Hz, 1H), 7.08 (d, J=8.9 Hz, 1H), 4.06 (c, J=7.2 Hz, 2H), 1.31(t, J=7.2 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 171.5, 169.5,164.1, 157.8, 137.8, 133.3, 126.4, 124.2, 115.1, 114.5, 97.5, 36.4,13.2. HPLC: Purity>99%, r.t.=8.3 min. ESI MS (m/z): 226 [M+H]⁺.

7-chloro-N-methyl isatoic anhydride (26).

Reagents: 4-chloro isatoic anhydride (4.55 mmols, 0.9 g); Sodium hydride(5.46 mmols, 0.9 g); Methyl iodide (13.7 mmols, 1.9 g). Yield: 0.4 g(42%), yellow solid, m.p.=215° C.-216° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 7.99 (d, J=8.4 Hz, 1H), 7.59 (d,J=1.8 Hz, 1H), 7.39 (dd, J=1.8, 8.4 Hz, 1H), 3.46 (s, 3H); ^(—)C NMR (75MHz, DMSO-d₆) δ (ppm): 158.6, 147.9, 143.9, 142.3, 131.4, 124.1, 115.4,111.1, 32.4. HPLC: Purity>96%, r.t.=7.3 min. ESI MS (m/z): 212 [M+H]⁺.

7-chloro-N-ethyl isatoic anhydride (27).

Reagents: 4-chloro isatoic anhydride (4.55 mmols, 0.9 g); Sodium hydride(5 mmols, 0.12 g); Ethyl iodide (8.3 mmols, 1.3 g). Yield: 0.65 g (63%),white solid, m.p.=156° C.-157° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 8.00 (d, J=8.4 Hz, 1H), 7.64 (d,J=1.7 Hz, 1H), 7.37 (dd, J=1.7, 8.4 Hz, 1H), 4.06 (c, J=7.0 Hz, 2H),1.20 (t, J=7.0 Hz, 3H); ^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 158.8,147.6, 142.8, 142.5, 131.8, 124.1, 114.9, 111.4, 40.2, 12.3. HPLC:Purity>97%, r.t.=8.1 min. ESI MS (m/z): 226 [M+H]⁺.

7-chloro-N-benzyl isatoic anhydride (28).

Reagents: 4-chloro isatoic anhydride (5 mmols, 1.0 g); Sodium hydride(7.83 mmols, 0.19 g); Benzyl bromide (10.1 mmols, 1.7 g). Yield: 0.9 g(38%), white solid, m.p.=207° C.-208° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 8.03 (d, J=8.3 Hz, 1H), 7.42 (m, 2H),7.37 (m, 1H), 7.35 (m, 3H), 7.29 (m, 1H), 5.31 (s, 2H); ^(—)C NMR (75MHz, DMSO-d₆) δ (ppm): 158.2, 148.1, 142.5, 141.5, 135.0, 131.2, 128.7,127.4, 126.6, 123.8, 114.9, 111.3, 47.6. HPLC: Purity>99%, r.t.=9.4 min.ESI MS (m/z): 288 [M+H]⁺.

General Synthesis Process for Compounds 29-41

Diethyl malonate is added drop by drop on a solution of sodium hydridein anhydrous DMF (15 ml) under a nitrogen atmosphere. The mixture isleft under stirring for 30 minutes at room temperature. Subsequently,the reaction is added to a flask containing the corresponding isatoicanhydride derivative dissolved in anhydrous DMF (15 ml) under a Nitrogenatmosphere. The mixture is heated to 50° C. for 5 hours. Dichloromethane(30 ml), water (50 ml) and an HCl solution (1 N) are added to themixture up to pH=5. The organic phase is separated, and the aqueousphase is once again extracted with dichloromethane (30 ml). The organicphases are brought together and washed with a saturated NaCl solution(30 ml). The organic phase is dried on anhydrous magnesium sulfate andfiltered, and the solvent is evaporated under vacuum. The resultingsolid is recrystallised in dichloromethane/hexane, to produce thedesired final product.

Ethyl 4-hydroxy-1-(isoprenyl)-2-oxo-1,2-dihydroquinoline-3-carboxylate(29)

Reagents: Comp 18 (1.3 mmols, 0.3 g); Sodium hydride (1.3 mmols, 0.065g); Diethyl malonate (6.5 mmols, 1.0 g). Yield: 0.08 g (21%), whitesolid, m.p.=123° C.-124° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 14.2 (s, 1H), 8.18 (dd, J=1.6, 8.3Hz, 1H), 7.65 (td, J=1.6, 7.1, 7.9 Hz, 1H), 7.24 (m, 2H), 5.10 (m, 1H),4.88 (d, J=6.0 Hz, 2H), 4.51 (c, J=7.1 Hz, 2H), 1.80 (s, 3H), 1.63 (s,3H), 1.41 (t, J=7.1 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 172.9,171.8, 159.5, 140.9, 135.8, 134.3, 125.9, 121.8, 119.7, 115.2, 114.7,98.0, 62.5, 40.9, 25.7, 18.5, 14.4. HPLC: Purity>96%, r.t.=10.2 min. ESIMS (m/z): 302 [M+H]⁺.

Ethyl 4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carboxylate(30).

Reagents: Comp 19 (1.36 mmols, 0.3 g); Sodium hydride (5.75 mmols, 0.138g); Diethyl malonate (6.84 mmols, 1.1 g). Yield: 0.15 g (38%), yellowoil.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 13.02 (s, 1H), 8.06 (dd, J=1.55, 8.3Hz, 1H), 7.72 (ddd, J=1.60, 7.0, 8.7 Hz, 1H), 7.55 (d, J=8.6 Hz, 1H),7.29 (ddd, J=0.86, 7.0, 7.9 Hz, 1H), 4.33 (c, J=8.6 Hz, 2H), 4.07 (d,J=7.4 Hz, 2H), 2.10 (sept, J=7.0 Hz, 1H), 1.31 (t, J=7.4 Hz, 3H), 0.89(d, J=7.4 Hz, 6H); ^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 169.2, 165.5,158.9, 140.0, 133.6, 124.6, 121.7, 115.3, 114.5, 101.0, 61.4, 47.5,26.6, 19.8, 13.9. HPLC: Purity>95%, r.t.=10.0 min. ESI MS (m/z): 290[M+H]⁺.

Ethyl1-(cyclopropylmethyl)-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate(31).

Reagents: Comp 20 (1.38 mmols, 0.3 g); Sodium hydride (2.91 mmols, 0.07g); Diethyl malonate (6.91 mmols, 1.1 g).

The reaction product (white solid) is not purified and is used for thefollowing reaction step. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 14.2 (s,1H), 8.13 (dt, J=1.25, 8.1 Hz, 1H), 7.61 (td, J=1.59, 7.27, 7.97 Hz,1H), 7.37 (d, J=8.63 Hz, 1H), 7.18 (m, 1H), 4.44 (c, J=7.12 Hz, 2H),4.12 (d, J=6.67 Hz, 2H), 1.41 (t, J=7.12 Hz, 3H), 1.25-1.10 (m, 1H),0.55-0.38 (m, 4H); HPLC: Purity>86%, r.t.=9.7 min. ESI MS (m/z): 288[M+H]⁺.

Ethyl 1-benzyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate (32).

Reagents: Comp 21 (3.4 mmols, 0.85 g); Sodium hydride (7.37 mmols, 0.18g); Diethyl malonate (16.8 mmols, 2.7 g). Yield: 0.1 g (9%), whitesolid, m.p.=117° C.-118° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 14.29 (s, 1H), 8.13 (dd, J=1.5, 8.0 Hz,1H), 7.48-7.43 (m, 1H), 7.23-7.18 (m, 2H), 7.15-7.11 (m, 5H), 5.44 (s,2H), 4.45 (c, J=7.1 Hz, 2H), 1.42 (t, J=7.1 Hz, 3H); ^(—)C NMR (75 MHz,CDCl₃) δ (ppm): 171.7, 171.0, 158.7, 139.8, 135.4, 133.3, 127.7, 126.2,125.4, 124.7, 121.0, 113.9, 96.8, 61.4, 44.6, 13.21. HPLC: Purity>99%,r.t.=9.9 min. ESI MS (m/z): 324 [M+H]⁺.

Ethyl 6-bromo-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate (33).

Reagents: 5-bromo isatoic anhydride (4.1 mmols, 1.0 g); Sodium hydride(8.3 mmols, 0.2 g); Diethyl malonate (20.8 mmols, 3.3 g).

The final product is not isolated and is used for the following reactionstep.

Ethyl 6-chloro-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate (34).

Reagents: 5-chloro isatoic anhydride (5.0 mmols, 1.0 g); Sodium hydride(10.4 mmols, 0.25 g); Diethyl malonate (25 mmols, 4.0 g).

The final product is not isolated and is used for the following reactionstep.

Ethyl6-bromo-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate(35).

Reagents: Comp 22 (2.3 mmols, 0.6 g); Sodium hydride (4.8 mmols, 0.12g); Diethyl malonate (11.5 mmols, 1.8 g). Yield: 0.3 g (40%), whitesolid, m.p.=139° C.-140° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 14.14 (s, 1H), 8.28 (d, J=2.4 Hz, 1H),7.74 (dd, J=2.4, 9.0 Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 4.50 (c, J=7.1 Hz,2H), 3.62 (s, 3H), 1.48 (t, J=7.1 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ(ppm): 172.5, 170.6, 159.3, 140.3, 137.2, 128.2, 116.5, 116.0, 115.0,98.7, 62.7, 29.4, 14.3. HPLC: Purity>96%, r.t.=9.6 min. ESI MS (m/z):328 [M+H]⁺.

Ethyl 6-bromo-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate(36).

Reagents: Comp 23 (1.8 mmols, 0.5 g); Sodium hydride (3.7 mmols, 0.09g); Diethyl malonate (4.25 mmols, 1.5 g). Yield: 0.3 g (48%), whitesolid, m.p.=129° C.-130° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 14.14 (s, 1H), 8.23 (d, J=2.4 Hz, 1H),7.67 (dd, J=2.4, 9.0 Hz 1H), 7.13 (d, J=9.0 Hz, 1H), 4.44 (c, J=7.1 Hz,2H), 4.20 (c, J=7.1 Hz, 2H), 1.42 (t, J=0.5, 7.1 Hz, 3H), 1.24 (t,J=1.5, 7.2 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 172.4, 170.4,158.7, 139.2, 137.0, 128.7, 116.6, 115.7, 114.6, 98.5, 62.5, 37.4, 14.2,12.7. HPLC: Purity>99%, r.t.=10.6 min. ESI MS (m/z): 342 [M+H]⁺.

Ethyl6-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate(37).

Reagents: Comp 24 (3.5 mmols, 0.8 g); Sodium hydride (7.1 mmols, 0.18g); Diethyl malonate (18 mmols, 2.8 g). Yield: 0.3 g (30%), white solid,m.p.=138° C.-139° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 14.13 (s, 1H), 8.07 (d, J=2.5 Hz, 1H),7.55 (dd, J=2.5, 9.0 Hz 1H), 7.20 (d, J=2.0 Hz, 1H), 4.44 (c, J=7.1 Hz,2H), 3.57 (s, 3H), 1.41 (t, J=7.1 Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ(ppm): 172.5, 170.6, 159.4, 139.9, 134.4, 127.9, 125.15, 116.1, 115.7,98.7, 62.6, 29.5, 14.3. HPLC: Purity>96%, r.t.=9.4 min. ESI MS (m/z):282 [M+H]⁺.

Ethyl6-chloro-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate(38).

Reagents: Comp 25 (3.5 mmols, 0.8 g); Sodium hydride (7.4 mmols, 0.17g); Diethyl malonate (17.7 mmols, 2.8 g). Yield: 0.5 g (48%), whitesolid, m.p.=140° C.-141° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 14.14 (s, 1H), 8.09 (d, J=2.5 Hz, 1H),7.54 (dd, J=2.5, 9.1 Hz 1H), 7.19 (d, J=2.0 Hz, 1H), 4.44 (c, J=7.1 Hz,2H), 4.21 (c, J=7.1 Hz, 2H), 1.42 (t, J=7.1 Hz, 3H), 1.25 (t, J=7.1 Hz,3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 171.4, 169.5, 157.7, 137.8,133.3, 126.4, 124.2, 115.1, 114.5, 97.5, 61.2, 36.4, 14.3, 13.2. HPLC:Purity>99%, r.t.=9.9 min. ESI MS (m/z): 296 [M+H]⁺.

Ethyl7-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate(39).

Reagents: Comp 26 (2.05 mmols, 0.4 g); Sodium hydride (4.16 mmols, 0.1g); Diethyl malonate (10.2 mmols, 1.65 g). Yield: 0.28 g (49%), whitesolid, m.p.=153° C.-154° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 12.95 (s, 1H), 8.02 (d, J=8.6 Hz,1H), 7.59 (d, J=1.8 Hz, 1H), 7.33 (dd, J=1.9, 8.7 Hz, 1H), 4.32 (c,J=7.1 Hz, 2H), 3.52 (s, 3H), 1.30 (t, J=7.1 Hz, 3H); ^(—)C NMR (75 MHz,DMSO-d₆) δ (ppm): 168.9, 164.6, 159.2, 141.7, 138.9, 126.7, 122.5,115.1, 113.7, 102.3, 61.8, 49.1, 29.4, 14.4. HPLC: Purity>99%, r.t.=5.1min. ESI MS (m/z): 282 [M+H]⁺.

Ethyl7-chloro-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate(40).

Reagents: Comp 27 (2.6 mmols, 0.6 g); Sodium hydride (4.6 mmols, 0.11g); Diethyl malonate (11.5 mmols, 1.85 g). Yield: 0.46 g (62%), yellowsolid, m.p.=141° C.-142° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 13.00 (s, 1H), 8.03 (d, J=8.6 Hz,1H), 7.64 (d, J=1.8 Hz, 1H), 7.33 (dd, J=1.9, 8.6 Hz, 1H), 4.32 (c,J=7.1 Hz, 2H), 4.19 (c, J=7.1 Hz, 2H), 1.30 (t, J=7.1 Hz, 3H), 1.15 (t,J=7.1 Hz, 3H); ^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 169.0, 164.8, 158.6,140.7, 139.0, 127.0, 122.3, 114.6, 113.9, 102.0, 61.8, 37.1, 14.4, 13.0.HPLC: Purity>99%, r.t.=9.9 min. ESI MS (m/z): 296 [M+H]⁺.

Ethyl1-benzyl-7-chloro-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylate(41).

Reagents: Comp 28 (1.9 mmols, 0.5 g); Sodium hydride (3.96 mmols, 0.09g); Diethyl malonate (9.6 mmols, 1.55 g). Yield: 0.9 g (94%), whitesolid, m.p.=172° C.-173° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 13.17 (s, 1H), 8.07 (d, J=8.6 Hz,1H), 7.43 (d, J=1.8 Hz, 1H), 7.35-7.32 (m, 3H), 7.26-7.23 (m, 1H), 7.18(dd, J=1.3, 8.6 Hz, 2H), 5.47 (s, 2H), 4.34 (c, J=7.1 Hz, 2H), 1.31 (t,J=7.1 Hz, 3H); ^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 168.5, 164.8, 158.9,140.6, 138.7, 136.5, 128.7, 127.1, 126.6, 126.4, 122.3, 114.9, 113.8,101.6, 61.5, 44.5, 13.9. HPLC: Purity>99%, r.t.=10.8 min. ESI MS (m/z):358 [M+H]⁺.

General Synthesis Process for Compounds 42-62

The corresponding ethyl carboxylate derivative (compounds 29-41),jointly with the corresponding hydrazide in DMF (0.3 ml), are added in aflask. The mixture is heated to 160° C. for 3 min. The reaction isallowed to cool at room temperature and, subsequently, MeOH is added (5ml), to form a precipitate that is filtered and purified with MeOHwashings.

4-Hydroxy-1-isoprenyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(42).

Reagents: Comp 29 (0.66 mmols, 0.2 g); octanoyl hydrazine (0.73 mmols,0.12 g). Yield: 0.125 g (22%), white solid, m.p.=114° C.-115° C.

¹H NMR (300 MHz, CDCl₃) δ (ppm): 15.37 (s, 1H), 12.38 (d, J=5.3 Hz, 1H),8.67-8.55 (m, 1H), 8.06 (d, J=7.9 Hz, 1H), 7.57 (t, J=7.9 Hz, 1H),7.22-7.14 (m, 2H), 5.1-5.02 (m, 1H), 4.81-4.76 (m, 2H), 2.26 (t, J=7.5Hz, 2H), 1.80 (s, 3H).1.7-1.52 (m, 5H), 1.38-1.1 (m, 8H), 0.79 (t, J=6.4Hz, 3H); ^(—)C NMR (75 MHz, CDCl₃) δ (ppm): 170.1, 169.6, 166.9, 161.7,139.7, 136.5, 134.09, 125.1, 122.4, 119.1, 115.8, 114.9, 96.4, 40.9,34.4, 31.8, 29.3, 29.1, 25.7, 26.6, 22.7, 18.5, 14.2. HPLC: Purity>99%,r.t.=13.7 min. ESI MS (m/z): 414 [M+H]⁺. Elementary Analysis(C₂₃H₁₃N₃O₄) Calculated: C, 66.81%; H, 7.51%; N, 10.16%. Found: C,66.73%; H, 7.74%; N, 10.25%.

N′-Dodecanoyl-4-hydroxy-1-isoprenyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(43).

Reagents: Comp 29 (0.66 mmols, 0.2 g); dodecanoyl hydrazine (0.73 mmols,0.16 g). Yield: 0.125 g (40%), white solid, m.p.=107° C.-108° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.28 (s, 1H), 11.97 (d, J=3.9 Hz,1H), 10.77 (d, J=3.9 Hz, 1H), 8.11 (dd, J=7.9, 1.5 Hz, 1H), 7.83 (td,J=9.18, 7.2, 1.6 Hz, 1H), 7.51 (d, J=8.6 Hz, 1H), 7.39 (t, J=8.6 Hz,1H), 5.51-5.05 (m, 1H), 4.95-4.85 (m, 2H), 2.23 (t, J=7.3 Hz, 2H), 1.85(s, 3H), 1.67 (s, 3H), 1.6-1.40 (m, 2H), 1.35-1.1 (m, 16H), 0.84 (t,J=6.5 Hz, 3H); ^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.7, 169.6, 167.3,160.9, 139.0, 135.8, 134.7, 124.7, 122.7, 119.2, 115.5, 114.9, 95.7,32.9, 31.3, 29.0, 28.9, 28.8, 28.7, 28.5, 25.3, 24.9, 22.1, 18.2, 13.9.HPLC: Purity>99%, r.t.=4.9 min. ESI MS (m/z): 470 [M+2H]⁺. ElementaryAnalysis (C₂₇H₃₇N₃O₄) Calculated: C, 69.35%; H, 7.98%; N, 8.99%. Found:C, 69.48%; H, 8.01%; N, 8.86%.

N′-Dodecanoyl-4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(44).

Reagents: Comp 30 (0.52 mmols, 0.15 g); dodecanoyl hydrazine (0.78mmols, 0.17 g). Yield: 0.05 g (21%), white solid, m.p.=112° C.-113° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.32 (s, 1H), 12.05 (d, J=4.1 Hz,1H), 10.79 (d, J=4.1 Hz, 1H), 8.12 (d, J=1.6 Hz, 1H), 7.9-7.75 (m, 1H),7.70 (d, J=8.5 Hz, 1H), 7.35 (t, J=7.6 Hz, 1H), 4.18 (d, J=7.5 Hz, 2H),2.30-2.05 (m, 3H), 1.52-1.35 (m, 2H), 1.25-1.05 (m, 16H), 0.95-0.80 (m,9H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.6, 169.4, 167.0, 161.5,139.4, 134.5, 124.6, 122.6, 115.9, 114.9, 95.5, 47.85, 31.3, 29.0, 28.9,28.7, 28.6, 28.5, 26.7, 24.9, 22.0, 19.8, 13.9. HPLC: Purity>97%,r.t.=6.6 min. ESI MS (m/z): 458 [M+H]⁺. Elementary Analysis (C₂₆H₃₉N₃O₄)Calculated: C, 68.24%; H, 8.59%; N, 9.18%. Found: C, 68.33%; H, 8.71%;N, 9.44%.

1-(cyclopropylmethyl)-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(45).

Reagents: Comp 31 (0.45 mmols, 0.13 g); dodecanoyl hydrazine (0.68mmols, 0.15 g). Yield: 0.09 g (49%), white solid, m.p.=121° C.-122° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.30 (s, 1H), 12.01 (d, J=4.1 Hz,1H), 10.76 (d, J=4.1 Hz, 1H), 8.12 (d, J=7.9 Hz, 1H), 7.8-7.6 (m, 2H),7.35 (t, J=7.6 Hz, 1H), 4.23 (d, J=6.9 Hz, 2H), 2.23 (t, J=7.6 Hz, 2H),1.6-1.42 (m, 2H), 1.32-1.17 (m, 16H), 0.84 (t, J=6.9 Hz, 3H), 0.52-0.42(m, 5H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.7, 169.5, 167.0, 161.3,139.3, 134.6, 124.6, 122.7, 115.8, 114.9, 95.6, 45.2, 32.9, 31.3, 29.0,28.9, 28.8, 28.7, 28.5, 25.0, 22.1, 13.9, 9.8, 3.7. HPLC: Purity>99%,r.t.=15.8 min. ESI MS (m/z): 456 [M+H]⁺. Elementary Analysis(C₂₆H₃₇N₃O₄) Calculated: C, 68.54%; H, 8.19%; N, 9.22%. Found: C,68.50%; H, 8.28%; N, 9.04%.

1-Benzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(46).

Reagents: Comp 32 (0.12 mmols, 0.04 g); dodecanoyl hydrazine (0.15mmols, 0.033 g). Yield: 0.045 g (75%), white solid, m.p.=150° C.-151° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 15.63 (s, 1H), 12.13 (d, J=4.1 Hz,1H), 8.25-8.08 (m, 2H), 7.3-7.1 (m, 7H), 5.46 (s, 2H), 2.35-2.10 (m,2H), 1.75-1.50 (m, 2H), 1.5-1.1 (m, 16H), 0.9-0.75 (m, 3H); ¹³C NMR (75MHz, DMSO-d₆) δ (ppm): 170.5, 168.2, 165.8, 161.1, 138.7, 134.9, 133.2,127.9, 126.4, 125.4, 121.6, 114.9, 114.2, 95.1, 44.9, 33.4, 30.9, 28.6,28.4, 28.3, 28.2, 24.4, 21.7, 13.1. HPLC: Purity>99%, r.t.=9.6 min. ESIMS (m/z): 493 [M+H]⁺. Elementary Analysis (C₂₉H₃₇N₃O₄) Calculated: C,70.85%; H, 7.59%; N, 8.55%. Found: C, 71.02%; H, 7.70%; N, 8.68%.

6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinol ine-3-carbohydrazide(47).

Reagents: Comp 33 (0.25 mmols, 0.07 g); dodecanoyl hydrazine (0.27mmols, 0.06 g). Yield: 0.103 g (85%), white solid, m.p.=286° C.-287° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.20 (s, 1H), 12.17 (sa, 1H), 11.9(d, J=4.0 Hz, 1H), 10.76 (d, J=4.1 Hz, 1H), 8.1-8.0 (m, 1H), 7.86 (dd,J=8.7 Hz, 1H), 7.33 (d, J=8.9 Hz, 1H), 2.2 (t, J=7.3 Hz, 2H), 1.60-1.42(m, 2H), 1.2 (s, 16H), 0.84 (t, J=6.6 Hz, 3H); ^(—)C NMR (75 MHz,DMSO-d₆) δ (ppm): 170.4, 169.4, 167.0, 161.5, 137.6, 136.0, 134.9,132.3, 126.0, 125.4, 117.9, 113.8, 32.6, 30.7, 28.4, 28.3, 28.2, 28.1,28.0, 24.4, 21.4, 13.2. ESI MS (m/z): 481. Elementary Analysis(C₂₂H₃₀BrN₃O₄) Calculated: C, 55.0%; H, 6.29%; N, 8.75%. Found: C,54.98%; H, 6.57%; N, 8.90%.

6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide(48).

Reagents: Comp 34 (0.14 mmols, 0.037 g); dodecanoyl hydrazine (0.15mmols, 0.034 g). Yield: 0.045 g (73%), white solid, m.p.=285° C.-286° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.22 (s, 1H), 12.18 (s, 1H), 11.9(d, J=4.2 Hz, 1H), 10.75 (sa, 1H), 7.97-7.90 (m, 1H), 7.76 (dd, J=2.46,8.8 Hz, 1H), 7.40 (d, J=8.8 Hz, 1H), 2.2 (t, J=7.3 Hz, 2H), 1.60-1.45(m, 2H), 1.2 (s, 16H), 0.85 (t, J=6.4 Hz, 3H).

6-Bromo-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(49).

Reagents: Comp 35 (0.46 mmols, 0.15 g); octanoyl hydrazine (0.5 mmols,0.08 g). Yield: 0.13 g (64%), white solid, m.p.=185° C.-186° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.30 (s, 1H), 11.94 (d, J=4.2 Hz,1H), 10.8 (d, J=4.2 Hz, 1H), 8.12 (s, 1H), 7.95 (d, J=9.3 Hz, 1H), 7.61(d, J=9.0 Hz, 1H), 3.6 (s, 3H), 2.23 (t, J=7.4 Hz, 2H), 1.6-1.42 (m,2H), 1.27 (s, 8H), 0.86 (t, J=6.2 Hz, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ(ppm): 169.4, 169.3, 166.8, 160.8, 138.7, 136.4, 126.0, 117.7, 116.2,114.5, 96.3, 32.7, 30.8, 29.2, 28.2, 27.9, 24.6, 21.6, 13.5. HPLC:Purity>98%, r.t.=9.8 min. ESI MS (m/z): 440 [M+H]⁺. Elementary Analysis(C₁₉H₂₄BrN₃O₄) Calculated: C, 52.0%; H, 5.52%; N, 9.59%. Found: C,52.06%; H, 5.50%; N, 9.77%.

6-Bromo-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(50).

Reagents: Comp 35 (0.36 mmols, 0.12 g); dodecanoyl hydrazine (0.4 mmols,0.08 g). Yield: 0.1 g (55%), white solid, m.p.=160° C.-161° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.23 (s, 1H), 11.85 (sa, J=4.3 Hz,1H), 10.5 (sa, J=5.5 Hz, 1H), 8.14 (d, J=2.2 Hz, 1H), 7.91 (dd, J=9.1,1.4 Hz, 1H), 7.58 (d, J=9.1 Hz, 1H), 3.6 (s, 3H), 2.26 (t, J=7.4 Hz,2H), 1.6-1.45 (m, 2H), 1.27 (s, 16H), 0.87 (t, J=6.4 Hz, 3H); ^(—)C NMR(75 MHz, DMSO-d₆) δ (ppm): 170.4, 167.8, 161.8, 139.7, 137.4, 127.0,118.6, 117.3, 115.4, 97.2, 32.6, 30.7, 28.9, 28.5, 28.4, 28.2, 28.0,24.4, 21.5, 13.2. HPLC: Purity>98%, r.t.=14.3 min. ESI MS (m/z): 494[M+H]⁺. Elementary Analysis (C₂₃H₃₂BrN₃O₄) Calculated: C, 55.8%; H,6.52%; N, 8.50%. Found: C, 55.94%; H, 6.80%; N, 8.61%.

6-Bromo-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(51).

Reagents: Comp 36 (0.36 mmols, 0.12 g); octanoyl hydrazine (0.4 mmols,0.06 g). Yield: 0.13 g (80%), white solid, m.p.=171° C.-172° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.24 (s, 1H), 11.88 (d, J=4.0 Hz.1H), 10.6 (d, J=4.1 Hz, 1H), 8.12 (d, J=2.5 Hz, 1H), 7.90 (dd, J=9.1,2.5 Hz, 1H), 7.61 (d, J=9.1 Hz, 1H), 4.3 (c, J=7.0 Hz, 2H), 2.22 (t,J=7.4 Hz, 2H), 1.57-1.50 (m, 2H), 1.30-1.17 (m, 11H), 0.84 (t, J=6.7 Hz,3H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 169.4, 166.7, 160.4, 137.8,136.7, 126.3, 117.5, 116.6, 114.4, 96.2, 36.9, 32.8, 30.9, 28.3, 28.1,24.7, 21.7, 13.6, 12.4. HPLC: Purity>98%, r.t.=13.5 min. ESI MS (m/z):452 [M+H]⁺.

6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(52).

Reagents: Comp 36 (0.36 mmols, 0.12 g); dodecanoyl hydrazine (0.4 mmols,0.08 g). Yield: 0.15 g (80%), white solid, m.p.=146° C.-147° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.28 (s, 1H), 11.97 (s, 1H), 10.8(s, 1H), 8.16 (s, 1H), 7.97 (dd, J=9.6, 2.5 Hz, 1H), 7.62 (d, J=8.8 Hz,1H), 4.3 (c, J=6.9 Hz, 2H), 2.24 (t, J=7.2 Hz, 2H), 1.6-1.45 (m, 2H),1.26 (s, 19H), 0.85 (t, J=8.6 Hz, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ(ppm): 169.3, 166.7, 160.3, 137.6, 136.4, 126.2, 117.2, 116.5, 114.2,96.0, 36.7, 32.6, 30.8, 28.5, 28.4, 28.2, 28.1, 28.0, 24.4, 21.5, 13.2,12.1. HPLC: Purity>98%, r.t.=6.2 min. ESI MS (m/z): 510 [M+2H]⁺.Elementary Analysis (C₂₄H₃₄BrN₃O₄) Calculated: C, 56.7%; H, 6.74%; N,8.26%. Found: C, 56.75%; H, 7.01%; N, 8.43%.

6-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(53).

Reagents: Comp 37 (0.36 mmols, 0.1 g); octanoyl hydrazine (0.39 mmols,0.062 g). Yield: 0.12 g (86%), white solid, m.p.=187° C.-188° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.33 (s, 1H), 11.96 (d, J=4.2 Hz,1H), 10.8 (d, J=4.2 Hz, 1H), 8.06 (d, J=2.6 Hz, 1H), 7.86 (dd, J=9.0,2.5 Hz, 1H), 7.68 (d, J=9.1 Hz, 1H), 3.6 (s, 3H), 2.24 (t, J=7.4 Hz,2H), 1.6-1.45 (m, 2H), 1.26 (s, 16H), 0.87 (t, J=6.2 Hz, 3H); ^(—)C NMR(75 MHz, DMSO-d₆) δ (ppm): 169.4, 166.8, 161.1, 160.8, 138.4, 133.7,126.9, 122.9, 117.5, 115.9, 96.2, 32.7, 30.8, 29.2, 28.2, 28.0, 24.6,21.6, 13.5. HPLC: Purity>97%, r.t.=12.3 min. ESI MS (m/z): 394 [M+H]⁺.Elementary Analysis (C₁₉H₂₄ClN₃O₄) Calculated: C, 57.9%; H, 6.14%; N,10.67%. Found: C, 58.10%; H, 6.27%; N, 10.95%.

6-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(54).

Reagents: Comp 37 (0.36 mmols, 0.10 g); dodecanoyl hydrazine (0.39mmols, 0.08 g). Yield: 0.143 g (90%), white solid, m.p.=156.5° C.-157.5°C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.33 (s, 1H), 11.96 (d, J=4.2 Hz,1H), 10.8 (d, J=4.2 Hz, 1H), 8.00 (d, J=2.5 Hz, 1H), 7.86 (dd, J=9.1,2.5 Hz, 1H), 7.69 (d, J=9.1 Hz, 1H), 3.6 (s, 3H), 2.23 (t, J=7.3 Hz,2H), 1.6-1.47 (m, 2H), 1.25 (s, 16H), 0.85 (t, J=6.4 Hz, 3H); ¹³C NMR(75 MHz, DMSO-d₆) δ (ppm): 169.6, 169.4, 166.8, 160.8, 138.4, 133.7,126.9, 122.9, 117.5, 115.8, 96.2, 32.9, 29.2, 28.6, 28.5, 28.3, 28.3,28.2, 24.6, 21.7, 13.5. HPLC: Purity>99%, r.t.=13.9 min. ESI MS (m/z):450 [M+H]⁺. Elementary Analysis (C₂₃H₃₂ClN₃O₄) Calculated: C, 61.39%; H,7.17%; N, 9.34%. Found: C, 61.46%; H, 7.44%; N, 9.60%.

6-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(55).

Reagents: Comp 38 (0.68 mmols, 0.2 g); octanoyl hydrazine (0.74 mmols,0.119 g). Yield: 0.19 g (70%), white solid, m.p.=160° C.-161° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.33 (s, 1H), 11.98 (d, J=4.1 Hz,1H), 10.8 (d, J=4.1 Hz, 1H), 8.03 (d, J=2.3 Hz, 1H), 7.86 (dd, J=9.2,2.6 Hz, 1H), 7.75 (d, J=9.2 Hz, 1H), 4.3 (c, J=6.9 Hz, 2H), 2.24 (t,J=7.3 Hz, 2H), 1.60-1.53 (m, 2H), 1.35-1.18 (m, 11H), 0.87 (t, J=6.4 Hz,3H); ^(—)C NMR (100 MHz, DMSO-d₆) δ (ppm): 170.3, 166.6, 161.3, 138.2,134.8, 127.7, 124.2, 118.2, 117.0, 97.1, 39.8, 37.8, 33.6, 31.7, 29.2,28.9, 25.6, 22.6, 14.5, 13.3. HPLC: Purity>98%, r.t.=13.1 min. ESI MS(m/z): 408 [M+H]⁺. Elementary Analysis (C₂₀H₂₆ClN₃O₄) Calculated: C,58.89%; H, 6.42%; N, 10.30%. Found: C, 59.11%; H, 6.38%; N, 10.50%.

6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(56).

Reagents: Comp 36 (0.68 mmols, 0.2 g); dodecanoyl hydrazine (0.75 mmols,0.16 g). Yield: 0.234 g (74%), white solid, m.p.=149° C.-150° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.3 (s, 1H), 11.9 (d, J=4.2 Hz, 1H),10.8 (d, J=4.2 Hz, 1H), 8.05 (d, J=2.6 Hz, 1H), 7.97 (dd, J=9.1, 2.6 Hz,1H), 7.62 (d, J=9.2 Hz, 1H), 4.3 (c, J=6.9 Hz, 2H), 2.23 (t, J=7.3 Hz,2H), 1.35-1.17 (m, 19H), 0.86 (t, J=7.4 Hz, 3H); ¹³C NMR (75 MHz,DMSO-d₆) δ (ppm): 169.2, 166.6, 160.3, 137.2, 133.6, 126.6, 123.1,117.0, 116.0, 96.1, 36.8, 32.6, 30.8, 28.5, 28.5, 28.4, 28.2, 28.1,28.0, 24.5, 21.5, 13.2, 12.1. HPLC: Purity>98%, r.t.=14.9 min. ESI MS(m/z): 464 [M+H]⁺. Elementary Analysis (C₂₄H₃₄ClN₃O₄) Calculated: C,62.12%; H, 7.39%; N, 9.06%. Found: C, 61.94%; H, 7.61%; N, 9.18%.

7-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(57).

Reagents: Comp 39 (0.36 mmols, 0.1 g); octanoyl hydrazine (0.39 mmols,0.063 g). Yield: 0.07 g (50%), white solid, m.p.=164° C.-165° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.31 (s, 1H), 11.89 (sa, 1H), 10.5(sa, 1H), 8.09 (dd, J=8.6, 2.5 Hz, 1H), 7.72-7.70 (m, 1H), 7.43-7.40 (m,1H), 3.64 (s, 3H), 2.25 (t, J=7.4 Hz, 2H), 1.6-1.42 (m, 2H), 1.29 (s,8H), 0.88 (t, J=6.6 Hz, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.1,169.5, 167.0, 161.1, 140.6, 139.2, 126.0, 122.6, 115.0, 113.5, 95.7,32.7, 30.8, 29.2, 29.1, 28.2, 28.0, 24.6, 21.7, 13.5. HPLC: Purity>99%,r.t.=7.5 min. ESI MS (m/z): 394 [M+H]⁺. Elementary Analysis(C₁₆H₂₄ClN₃O₄) Calculated: C, 57.94%; H, 6.14%; N, 10.67%. Found: C,57.85%; H, 6.42%; N, 10.88%.

7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(58).

Reagents: Comp 39 (0.36 mmols, 0.10 g); dodecanoyl hydrazine (0.39mmols, 0.08 g). Yield: 0.105 g (65%), white solid, m.p.=165° C.-166° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.30 (s, 1H), 11.89 (sa, 1H), 10.8(sa, 1H), 8.06 (d, J=8.6 Hz, 1H), 7.74 (d, J=1.6, Hz, 1H), 7.69 (dd,J=8.6, 1.7 Hz, 1H), 3.6 (s, 3H), 2.23 (t, J=7.2 Hz, 2H), 1.53-1.27 (m,2H), 1.23 (s, 16H), 0.84 (t, J=6.5 Hz, 3H); ^(—)C NMR (75 MHz, DMSO-d₆)δ (ppm): 170.0, 169.4, 166.9, 161.0, 140.6, 139.1, 126.0, 122.6, 114.9,113.5, 113.4, 32.7, 30.9, 29.2, 29.1, 28.6, 28.5, 28.3, 28.3, 28.2,24.6, 21.7, 13.5. HPLC: Purity>98%, r.t.=13.8 min. ESI MS (m/z): 450[M+H]⁺. Elementary Analysis (C₂₃H₃₂ClN₃O₄) Calculated: C, 61.39%; H,7.17%; N, 9.34%. Found: C, 61.12%; H, 7.19%; N, 9.61%.

7-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(59).

Reagents: Comp 40 (0.5 mmols, 0.15 g); octanoyl hydrazine (0.55 mmols,0.09 g). Yield: 0.06 g (29%), white solid, m.p.=161° C.-162° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.31 (s, 1H), 11.91 (d, J=4.1 Hz,1H), 10.8 (d, J=4.1 Hz, 1H), 8.09 (d, J=8.6 Hz, 1H), 7.90 (d, J=1.9 Hz,1H), 7.61 (dd, J=8.6, 1.8 Hz, 1H), 4.3 (c, J=7.0 Hz, 2H), 2.23 (t, J=7.3Hz, 2H), 1.60-1.45 (m, 2H), 1.35-1.12 (m, 11H), 0.86 (t, J=6.6 Hz, 3H);^(—)C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.0, 169.4, 166.9, 160.7, 139.5,139.3, 126.3, 122.6, 114.6, 113.7, 95.6, 36.9, 32.7, 30.8, 28.2, 28.0,24.6, 21.6, 13.5, 12.3. HPLC: Purity>99%, r.t.=8.2 min. ESI MS (m/z):408 [M+H]⁺. Elementary Analysis (C₂₀H₂₆ClN₃O₄) Calculated: C, 58.9%; H,6.42%; N, 10.30%. Found: C, 59.13%; H, 6.70%; N, 10.58%.

7-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(60).

Reagents: Comp 40 (0.5 mmols, 0.15 g); dodecanoyl hydrazine (0.56 mmols,0.124 g). Yield: 0.221 g (95%), white solid, m.p.=138° C.-139° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.17 (s, 1H), 11.91 (sa, 1H), 10.8(sa, 1H), 8.11 (d, J=8.6 Hz, 1H), 7.75 (d, J=1.8 Hz, 1H), 7.42 (dd,J=8.6, 1.8 Hz, 1H), 4.3 (c, J=6.9 Hz, 2H), 2.23 (t, J=7.2 Hz, 2H),1.54-1.40 (m, 2H), 1.37-1.10 (m, 19H), 0.84 (t, J=6.3 Hz, 3H); ^(—)C NMR(75 MHz, DMSO-d₆) δ (ppm): 169.9, 169.3, 166.7, 160.6, 139.4, 139.2,126.2, 122.4, 114.3, 113.7, 95.5, 36.7, 32.6, 30.8, 28.5, 28.4, 28.3,28.2, 24.5, 21.6, 13.3, 12.1. HPLC: Purity>98%, r.t.=14.7 min. ESI MS(m/z): 464 [M+H]⁺. Elementary Analysis (C₂₄H₃₄ClN₃O₄) Calculated: C,62.1%; H, 7.39%; N, 9.06%. Found: C, 62.33%; H, 7.57%; N, 9.24%.

1-Benzyl-7-chloro-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide (61).

Reagents: Comp 41 (0.5 mmols, 0.20 g); octanoyl hydrazine (0.62 mmols,0.100 g). Yield: 0.196 g (74%), white solid, m.p.=182° C.-183° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.51 (s, 1H), 11.86 (d, J=4.2 Hz,1H), 10.8 (d, J=4.2 Hz, 1H), 8.11 (d, J=8.6 Hz, 1H), 7.57 (d, J=1.8 Hz,1H), 7.42 (dd, J=8.6, 1.8 Hz, 1H), 7.36-7.30 (m, 2H), 7.28-7.20 (m, 3H),5.6 (s, 2H), 2.24 (t, J=7.4 Hz, 2H), 1.60-1.50 (m, 2H), 1.35-1.15 (m,8H), 0.86 (t, J=6.8 Hz, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.7,169.6, 166.9, 161.5, 140.1, 139.3, 136.1, 128.7, 127.3, 126.7, 126.4,123.2, 115.5, 114.1, 95.8, 44.8, 32.9, 31.2, 28.5, 28.4, 24.9, 22.1,13.9. HPLC: Purity>98%, r.t.=2.4 min. ESI MS (m/z): 470 [M+H]⁺.Elementary Analysis (C₂₅H₂₈ClN₃O₄) Calculated: C, 63.89%; H, 6.01%; N,8.94%. Found: C, 63.47%; H, 6.30%; N, 9.08%.

1-Benzyl-7-chloro-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(62).

Reagents: Comp 41 (0.56 mmols, 0.20 g); dodecanoyl hydrazine (0.62mmols, 0.133 g). Yield: 0.25 g (85%), white solid, m.p.=167° C.-168° C.

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 16.49 (s, 1H), 11.87 (d, J=4.2 Hz,1H), 10.87 (d, J=4.2 Hz, 1H), 8.11 (d, J=8.6 Hz, 1H), 7.57 (d, J=1.8 Hz,1H), 7.41 (dd, J=8.6, 1.8 Hz, 1H), 7.36-7.30 (m, 2H), 7.28-7.20 (m, 3H),5.6 (s, 2H), 2.23 (t, J=7.3 Hz, 2H), 1.60-1.50 (m, 2H), 1.30-1.10 (m,16H), 0.84 (t, J=6.8 Hz, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm): 170.7,169.5, 166.9, 161.5, 140.1, 139.3, 136.1, 128.7, 127.2, 126.6, 126.4,123.2, 115.5, 114.1, 95.8, 44.8, 32.9, 31.3, 29.0, 28.9, 28.7, 28.7,28.5, 24.9, 22.1, 13.9. HPLC: Purity>99%, r.t.=5.1 min. ESI MS (m/z):526 [M+H]⁺. Elementary Analysis (C₂₉H₃₆ClN₃O₄) Calculated: C, 66.21%; H,6.90%; N, 7.99%. Found: C, 66.21%; H, 7.06%; N, 8.12%.

Synthesis ofN′-1-dibenzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide(Comp 63)

A mixture containing benzyl bromide (1.88 mmols, 0.32 g), cesiumcarbonate (1.88 mmol, 0.363 g) and Compound 13 (1.24 mmol, 0.5 g) in DMF(2 ml) is microwave-heated for 1 h at 160° C.

Dichloromethane (25 ml) and water (25 ml) are added to the mixture. Theorganic phase is separated, and the aqueous phase is once againextracted with dichloromethane (25 ml). The organic phases are broughttogether and washed with a saturated NaCl solution (30 ml). The organicphase is dried on anhydrous magnesium sulfate and filtered, and thesolvent is evaporated under vacuum. The resulting solid is purified bymeans of silica gel plates using Ethyl acetate:hexane (1:20) as theeluents. A white solid is obtained, 11 mg (2%).

¹H NMR (300 MHz, DMSO-d₆) δ (ppm): 15.94 (s, 1H), 11.80 (s, 1H), 8.13(d, J=8.0 Hz, 1H), 7.73 (t, J=7.8 Hz, 1H), 7.48 (d, J=8.7 Hz, 1H),7.58-7.30 (m, 11H), 5.5 (s, 2H), 4.77 (s, 2H), 2.36 (t, J=7.4 Hz, 2H),1.70-1.52 (m, 2H), 1.43-1.2 (m, 16H), 0.81 (t, J=7.6 Hz, 3H); ^(—)C NMR(75 MHz, DMSO-d₆) δ (ppm): 173.9, 171.5, 170.6, 161.5, 139.4, 136.4,136.3, 134.9, 128.7, 128.5, 128.2, 127.4, 127.2, 126.5, 114.9, 96.0,50.45, 44.8, 31.5, 31.3, 29.0, 28.9, 28.7, 28.5, 24.2, 22.1, 14.0. HPLC:Purity>99%, r.t.=14.6 min. ESI MS (m/z): 582 [M+H]⁺.

Measurement of the Inhibition of GSK-3β

The enzyme inhibition assays were performed using the luminometricmethodology of kinase-glo®. The recombinant human GSK3 enzyme (catalogueno. 14-306) was acquired from Upstate (Dundee, UK). Theprephosphorylated polypeptide was synthesised by American Peptide Inc.(Sunnyvale, Calif.). The luminescent kinase kit (catalogue no. V6711)was obtained from Promega. The ATP and the other reagents were purchasedfrom Sigma-Aldrich (St. Louis, Mo.).

The assays were performed in buffer using 96-well plates. 10 μl of thecompound to be assayed (dissolved in DMSO at a concentration of 1 mM,and, in turn, dissolved in buffer to the concentration necessary for theexperiment) and 10 μl (20 ng) of the enzyme are added to each well,followed by 20 μl of buffer containing 25 μM of the substrate and 1 μMof ATP. The final concentration of DMSO in the experiment did not exceed1%. Following a half-hour incubation at 30° C., the enzymatic reactionis stopped with 40 μl of the kinase-glo® reagent. The luminescence ismeasured after ten minutes, using a POLARstar Optima multimode reader.The activity is proportional to the difference between the total ATP andthe ATP consumed. The inhibitory activities were calculated as afunction of the maximum activity, measured in the absence of inhibitor.

TABLE 1 Inhibitory concentration 50 (IC50) of some of the compounds withthe formula (I) Compound IC50 GSK-3β (μM) 3 8.67 ± 0.40 4 7.35 ± 0.23 62.85 ± 0.36 13 4.47 ± 0.29 14 9.00 ± 0.30 15 3.12 ± 0.03 17 5.81 ± 0.7642 7.11 ± 0.25 43 7.78 ± 0.15 45 5.25 ± 0.58 46 5.51 ± 0.27 47 3.54 ±0.18 48 3.81 ± 0.12 49 6.28 ± 0.33 50 4.28 ± 0.50 51 7.34 ± 0.20 52 2.01± 0.18 53 8.48 ± 0.30 54 5.95 ± 0.49 55 9.42 ± 0.23 56 3.18 ± 0.28 576.66 ± 0.29 58 4.03 ± 0.19 59 4.83 ± 0.23 60 2.48 ± 0.26 61 5.96 ± 0.1762 5.99 ± 0.22 63 6.84 ± 0.51

Kinetic Studies on the Inhibition of GSK-3β

In order to determine the binding mode of these compounds, differentkinetic studies were performed, varying the reaction times and the ATPconcentration and the substrate concentration. Thus, when the kineticstudy is performed at different enzyme incubation times with inhibitor 6and at two different inhibitor concentrations, it may be observed thatthe % of enzyme inhibition does not vary; therefore, we may concludethat it is a reversible inhibitor.

TABLE 2 Study of the inhibition of GSK-3β by compound 6 as a function oftime. Pre- incubation time [Comp. 6] 10 μM [Comp. 6] 5 μM E + I (min) %inhibition deviation % inhibition deviation 0 61.400072 3.6893298727.4181566 0.94012209 5 63.0369001 5.63307429 30.3138744 4.02663481 1062.2631193 1.73621051 30.1825742 1.13957388

The kinetic studies conducted by varying the ATP concentration from 1 to50 μM, and the concentration of inhibitor 6 from 5 to 10 μM, wereperformed using the ADP-Glo™ Kinase Assay. The double-reciprocal plot(FIG. 1) shows that the intercept on the vertical axis (1V) increaseswhen the concentration of compound 6 increases (from 5 to 10 μM),whereas the intercept on the horizontal axis (1[ATP]) does not change.This means that, whereas the maximum enzyme activity (Vmax) decreases inthe presence of the inhibitor, the Michaelis-Menten constant (Km)remains invariable. These results are compatible with the fact that 6acts as a non-ATP-competitive inhibitor of GSK-3β, since an increase inthe ATP concentration (from 1 to 50 μM) does not interfere with theenzyme inhibition.

On the other hand, we also studied the dependence of GSK-3β activity inthe presence of these inhibitors as a function of the concentration ofthe substrate, the GS-2 peptide, used in the assays. In these kineticstudies, the GS-2 concentration was varied from 15.5 to 100 μM and theATP concentration was kept constant at 1 μM. The double-reciprocal plot(FIG. 2) once again shows that the intercept on the vertical axis (1V)increases when the concentration of compound 6 increases (from 5 to 10μM), whereas the intercept on the horizontal axis (1[GS-2]) does notchange. This means that, whereas the maximum enzyme activity (Vmax)decreases in the presence of the inhibitor, the Michaelis-Mentenconstant (Km) remains invariable. These results are compatible with thefact that compound 6 acts as a substrate-non-competitive inhibitor ofGSK-3β, since an increase in the GS-2 concentration (from 15.5 to 100μM) does not interfere with the enzyme inhibition.

All the enzymatic studies, both the activity level studies and thekinetic studies, described herein are compatible with allostericmodulation of the GSK-3β enzyme by these inhibitors.

Determination of the Anti-Inflammatory Activity in Cellular Models

A monolayer culture of RAW 264.7 cells growing in 96-well plates isused. The cells are treated for 1 h with the compounds evaluated.Subsequently, the inflammatory activity is stimulated with 0.4 μg/ml ofLPS for 24 h. Once this time has elapsed, the production of nitrites isevaluated by means of the Griess method. Briefly, 50 μl of supernatantare transferred to 96-well plates and 50 μl of Griess reagent are added.The mixture is incubated for 15 minutes at room temperature and theabsorbance at 520 nm is measured in a microplate reader.

In order to determine the capacity of the allosteric GSK-3β inhibitorsto reduce the activation of inflammation in cell cultures, compound 6was evaluated at various concentrations (1.25, 2.5, 5 and 10 μM), and adose-dependent effect on the decrease in nitrite release was found.

1. A method of treatment and/or prevention of a disease selected fromneurodegenerative diseases, inflammatory diseases, cancer or diabetescomprising the administration of an effective amount of a compound withthe formula (I)

or the salts, solvates or stereoisomers thereof, where R₁ is selectedfrom H, an optionally substituted C₁-C₅ alkyl group or an optionallysubstituted C₂-C₅ alkenyl group, R₂ is an optionally substituted C₅-C₁₅alkyl group, R₃ is selected from H, halogen, an optionally substitutedC₁-C₅ alkyl group or an optionally substituted C₁-C₅ —(O)-alkyl group, nis a value between 1 and 4, R₄, R₅ and R₆ are independently selectedfrom H or an optionally substituted C₁-C₅ alkyl group, provided that,when R₁ is ethyl or H and R₃, R₄, R₅ and R₆ are H, R₂ cannot be heptyl,2. The method for use according to claim 1, where R₄, R₅ and R₆ are H.3. The method according to claim 1, where R₁ is selected from H,optionally substituted methyl, ethyl or isobutyl or C₄ alkenyl,preferably isoprenyl.
 4. The method according to claim 3, where R₁ ismethyl substituted by cyclopropyl or phenyl.
 5. The method according toclaim 1, where R₂ is selected from a C₇ alkyl group or a C₁₁ alkylgroup.
 6. The method according to claim 1, where R₃ is selected from H,Br, Cl or F.
 7. The method according to claim 1, where n is
 1. 8. Themethod according to claim 1, where said compound is selected from thefollowing group:4-Hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide7-Chloro-4-hydroxy-N′-octanoyl-2-oxo-2-hydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Fluoro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide4-Hydroxy-1-(isoprenyl)-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-1-(isoprenyl)-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-(Cyclopropylmethyl)-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-4-hydroxy-1-methyl-N-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinone-3-carbohydrazide7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-7-chloro-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-7-chloro-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′,1-Dibenzyl-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide9. The method according to claim 1, selected from the following group:N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide10. The method according to claim 1, where the neurodegenerative diseaseis selected from Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis, cerebral ischaemia, post-encephalitic Parkinsonisms,dystonias, Tourette syndrome, periodic limb movement pathologies,restless legs syndrome, attention deficit hyperactivity disorders,Huntington's disease, progressive supranuclear palsy, Pick's disease,fronto-temporal dementia or neuromuscular diseases; where theinflammatory disease is selected from Crohn's disease, ulcerativecolitis, rheumatoid arthritis, atherosclerosis, vasculitis or multiplesclerosis; where the cancer is selected from glioblastoma, leukaemias,lymphomas, or lung, breast, prostate or colon cancer; and where thediabetes is non-insulin-dependent type II diabetes.
 11. Compound withthe formula (II)

or the salts, solvates or stereoisomers thereof, where R₁ is selectedfrom H, an optionally substituted C₁-C₅ alkyl group or an optionallysubstituted C₂-C₅ alkenyl group, R₂ is an optionally substituted C₅-C₁₅alkyl group, R_(3 is a halogen), n is a value between 1 and 4, R₄, R₅and R₆ are independently selected from H or an optionally substitutedC₁-C₅ alkyl group.
 12. Compound according to claim 11, where R₄, R₅ andR₆ are H.
 13. Compound according to claim 11, where R₁ is selected fromH, optionally substituted methyl, ethyl or isobutyl, or C₅ alkenyl,preferably isoprenyl.
 14. Compound according to claim 13, where R₁ ismethyl substituted by cyclopropyl or phenyl.
 15. Compound according toclaims 11, where R₂ is selected from a C₁₁ alkyl group or a C₇ alkylgroup.
 16. Compound according to claims 11, where R₃ is selected frombromine, chlorine or fluorine.
 17. Compound according to claim 16, wheren is
 1. 18. Compound selected from the following group:7-Chloro-4-hydroxy-N′-octanoyl-2-oxo-2-hydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Fluoro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide4-Hydroxy-1-(isoprenyl)-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-1-(isoprenyl)-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′-Dodecanoyl-4-hydroxy-1-isobutyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-(Cyclopropylmethyl)-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-N′-dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-4-hydroxy-1-methyl-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-1-ethyl-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-7-chloro-4-hydroxy-N′-octanoyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide1-Benzyl-7-chloro-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazideN′,1-Dibenzyl-N′dodecanoyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide19. Compound selected from the following group:N′-Dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-4-hydroxy-2-oxo-2-hydroquinoline-3-carbohydrazide6-Bromo-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide6-Chloro-N′-dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N′-dodecanoyl-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carbohydrazide7-Chloro-N%dodecanoyl-1-ethyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carbohydrazide20. Pharmaceutical composition that comprises a compound according toclaim 11, which optionally comprises another active principle. 21-38.(canceled)